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In Vitro and in Cellulo Sensing of Transition Metals Using Time-Resolved Fluorescence Spectroscopy and Microscopy

Pal, Robert; Barker, Abigail C.J.; Hummel, Daniel; Pålsson, Lars-Olof

In Vitro and in Cellulo Sensing of Transition Metals Using Time-Resolved Fluorescence Spectroscopy and Microscopy Thumbnail


Abigail C.J. Barker

Daniel Hummel


In this work we demonstrate that time domain techniques can be used successfully to monitor realtively weak modulations of the fluorescence in sensing applications. The metal sensing complex Newport Green DCF™ can detect selected transition metals in vivo as well as in vitro. Incremental addition of Ni and/or Zn (in vitro) lead to a substantial reduction in the yield of the fast component in a bi-exponential fluorescence decay (τ1 = 150–250 ps) from 60% to 30–35%. This is rationalised as an inhibition of intra-molecular electron transfer in the NPG sensing complex due to metal complexation. In order to explore this effect in cellulo, NIH 3 T3 mouse skin fibroplast cells were pre-incubated with set levels of Ni and Zn, at a constant concentration of NPG. The fluorescence modulation in cellullo was subsequently studied employing both time-resolved fluorescence microscopy and confocal fluorescence microscopy. In correlation with the in vitro observations, similar effects were observed on the fluorescence decay in cellulo.

Journal Article Type Article
Acceptance Date Dec 11, 2018
Online Publication Date Dec 26, 2018
Publication Date Jan 31, 2019
Deposit Date Jan 4, 2019
Publicly Available Date Jan 4, 2019
Journal Journal of Fluorescence
Print ISSN 1053-0509
Electronic ISSN 1573-4994
Publisher Springer
Peer Reviewed Peer Reviewed
Volume 29
Issue 1
Pages 255-263
Public URL


Published Journal Article (Advance online version) (1.7 Mb)

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Copyright Statement
Advance online version © The Author(s) 2018.
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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